Novel nucleic acid-based tracers and their uses thereof

Abstract

The present invention relates to a method of determining whether a facility can effectively prevent the spread of an infectious disease by a) releasing an effective amount of biological tracers in or near the facility, to mimic the spread of pathogens that cause the infectious disease; and b) detecting the presence or amount of the biological tracers to determine the spread of the biological tracers in the environment of the facility. The present invention further provides a method and apparatus to generate aerosol that mimic the human sneeze and cough so as to more realistically determine the spread of an infectious disease by such a source.

1 . A method of determining whether a facility can effectively prevent the spread of an infectious disease comprising: 
 a) releasing an effective amount of biological tracers in or near the facility, to mimic the spread of pathogens that cause the infectious disease; and    b) detecting the presence or amount of the biological tracers to determine the spread of the biological tracers in the environment of the facility.    
     
     
         2 . The method of  claim 1  wherein the facility is used to prevent the spread of the infectious disease from the outside into the facility, the biological tracers are released outside the facility, and the presence or amount of the biological tracers is detected in the facility.  
     
     
         3 . The method of  claim 1  wherein the facility is used to prevent the spread of the infectious disease from the inside to the outside of the facility, the biological tracers are released in the facility, and the presence or amount of the biological tracers is detected outside the facility.  
     
     
         4 . The method of  claim 1  wherein the biological tracers are unique from any other biological molecules existing in the facility and its environment.  
     
     
         5 . The method of  claim 1  further comprising collecting a sample in or near the facility, the presence or amount of the biological tracers is determined by analyzing the sample.  
     
     
         6 . The method of  claim 5  wherein the sample is collected from the medium in or near the facility, wherein the medium is selected from the group consisting of gas, liquid and solid substance.  
     
     
         7 . The method of  claim 6  wherein the gas substance is air, to thereby determine whether the infectious disease has spread into the air in or near the facility.  
     
     
         8 . The method of  claim 6  wherein the liquid is water, to thereby determine whether the infectious disease has spread into the water in or near the facility.  
     
     
         9 . The method of  claim 6  wherein the solid substance is collected from the surface of a solid object, to thereby determine whether the biological tracer has landed on or adhered to the surface of the solid object.  
     
     
         10 . The method of  claim 1  wherein the biological tracers are nucleic acids.  
     
     
         11 . The method of  claim 10  wherein the nucleic acids are selected from the group consisting of single stranded DNAs, single stranded RNAs, double stranded DNAs, double stranded RNAs, DNA/RNA hybrids, chimeric DNA molecules, chimeric RNA molecules, plasmid DNAs, and DNAs and RNA extracted from an organism.  
     
     
         12 . The method of  claim 11  wherein the organism is selected from the group consisting of  E. coli , yeast, and other organisms.  
     
     
         13 . The method of  claim 10  wherein the nucleic acids are identified by a method comprising a step selected from the group consisting of nucleic acid hybridization, polymerase chain reaction, restriction fragment length polymorphism, and sequence information.  
     
     
         14 . The method of  claim 10  wherein each of the nucleic acids consists of any number of nucleotides.  
     
     
         15 . The method of  claim 10  wherein each of the nucleic acids consists of between 100-5,000 nucleotides.  
     
     
         16 . The method of  claim 10  wherein each of the nucleic acids consists of between 40-100 nucleotides.  
     
     
         17 . The method of  claim 1  wherein the detecting step comprising amplifying the biological tracers.  
     
     
         18 . The method of  claim 17  wherein nucleic acid amplification technology is used to amplify the biological tracers.  
     
     
         19 . The method of  claim 1  wherein the facility is selected from the group consisting of ventilation system and drainage system.  
     
     
         20 . The method of  claim 1  wherein the biological tracer is provided in a form selected from the group consisting of aerosol and solution.  
     
     
         21 . The method of  claim 20  wherein the solution of the biological tracer comprises an additive substance that increases the viscosity of the solution to more closely mimic the body fluids in which viruses are released into the environment.  
     
     
         22 . The method of  claim 21  wherein the substance that increases the viscosity of the solution is selected from the group consisting of proteins, gelatin, and glycerol.  
     
     
         23 . The method of  claim 20  wherein the infectious disease is an airborne disease, the solution is released in the facility in a range of fairly uniformly sized droplets, and the droplets have a diameter of equal to or less than 1 micrometer.  
     
     
         24 . The method of  claim 20  wherein the infectious disease is droplet-disseminated disease, the solution is released in the facility in a range of substantially uniformly sized droplets, and the droplets have a diameter of from 1 to 10 micrometer.  
     
     
         25 . The method of  claim 20  wherein the concentration of the solution ranges from 2×10 −15  to 1 mmole/l, preferably from 2×10 −15  to 2×10 −5  mmole/l, and even more preferably from 2×10 −15  to 2×10 −9  mmole/l.  
     
     
         26 . The method of  claim 20  wherein the concentration of the solution ranges from 10 3  to 5×10 17  copies/ml, preferably from 10 3  to 10 13  copies/ml, and even more preferably from 10 3  to 10 9  copies/ml.  
     
     
         27 . The method of  claim 1  wherein the biological tracer is not encapsulated by a polymer.  
     
     
         28 . The method of  claim 1  wherein the biological tracer is encapsulated by a polymer.  
     
     
         29 . The method of  claim 28  wherein the polymer is selected from the group consisting of protein molecules, lipid molecules, carbohydrates, and combination thereof.  
     
     
         30 . The method of  claim 1  wherein the sequences of the biological tracers encode information selected from the group consisting of manufacturer, manufacture date, sequence identification, lot number, and combinations thereof.  
     
     
         31 . The method of  claim 1  wherein the detection of the biological tracers at different locations at a given time after release of said tracers within the facility provides spatial information on the dispersal of said tracers.  
     
     
         32 . The method of  claim 1  wherein the detection of the biological tracers at different times after the release of said tracers at the same spot within the facility provides temporal information on the dispersal of said tracers.  
     
     
         33 . The method of  claim 1  wherein the detection of the biological tracers at different times and at different locations after the release of said tracers within the facility provides temporal and spatial information on the dispersal of said tracers.  
     
     
         34 . An apparatus for generating aerosol comprising a chamber of compressed gas having a nozzle that is attached to a detachable stiff tubing capable of holding a solution of predetermined volume.  
     
     
         35 . The apparatus of  claim 34  further comprising a trigger to open the nozzle so as to discharge the compressed gas through the tubing.  
     
     
         36 . The apparatus of  claim 34  wherein the chamber can be manually filled with compressed air, the pressure of which can be monitored by a pressure gauge.  
     
     
         37 . The apparatus of  claim 34  wherein the stiff tubing has a uniform internal luminal diameter.  
     
     
         38 . A method of generating aerosol that mimic the results of a human sneeze or cough comprising passing compressed air in pulses through a narrow tube that holds a solution being aerosolized.  
     
     
         39 . The method of  claim 38  wherein the repeated generation of pulses of aerosol simulate spells of coughing or sneezing.  
     
     
         40 . An apparatus that samples a body of air by suctioning into a vacuumed container through a temporary puncture or by manually drawing a predetermined volume of air into a chamber using a piston.  
     
     
         41 . The apparatus of  claim 40  wherein the vacuumed container or the chamber is sealed before being brought into the body of air being sampled.  
     
     
         42 . The apparatus of  claim 40  wherein the vacuumed container or the chamber contains a DNase-free electrolyte solution.  
     
     
         43 . The apparatus of  claim 40  wherein the container or the chamber is sealed after a sample of air is taken into the container or chamber.